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Abstract The remnants of Hurricane Ida caused major damage and death in the United States on September 1st, 2021, and 11 people drowned in flooded basement apartments within New York City (NYC). It was catastrophic because the maximum hourly precipitation intensity, recorded as 3.47 inches (88.1 mm) per hour at Central Park, was unprecedentedly high for the NYC region. The stormwater infrastructure in NYC is built for 1.75 inches (44.5 mm) per hour, and so understanding the dynamic risk associated with Ida can inform city planning efforts for climate change’s impact on short duration extreme precipitation events. We contextualize this storm’s record-breaking hourly intensity within the historical record as well as project its risk in the near- to medium-term future using nonstationary stochastic models. These models are conditioned on average temperature (T avg ) and cooling degree day (CDD) projections from three climate models as a covariate, each with a SSP 126 and SSP 370 scenario. The likelihood of such a storm was slowly increasing even before Ida happened, but the projected aggregate reoccurrence risk of an event of Ida’s magnitude over time from the non-stationary models ranges from 4 to 52 times higher than the risk given by the stationary model. Using CDD as a covariate resulted in risks that were more than twice the magnitude than when using T avg . Presenting both covariates provides a broader envelope of uncertainty, which highlights the importance and nuances in the choice of a regionally appropriate covariate for non-stationary risk analysis.

Abstract Background Prenatal alcohol exposure (PAE) has serious physical consequences for children such as behavioral disabilities, growth disorders, neuromuscular problems, impaired motor coordination, and decreased muscle tone. However, it is not known whether loss of muscle strength occurs, and which interventions will effectively mitigate physical PAE impairments. We aimed to investigate whether physical alteration persists during adolescence and whether exercise is an effective intervention. Results Using paradigms to evaluate different physical qualities, we described that early adolescent PAE animals have significant alterations in agility and strength, without alterations in balance and coordination compared to CTRL animals. We evaluated the effectiveness of 3 different exercise protocols for 4 weeks: Enrichment environment (EE), Endurance exercise (EEX), and Resistance exercise (REX). The enriched environment significantly improved the strength in the PAE group but not in the CTRL group whose strength parameters were maintained even during exercise. Resistance exercise showed the greatest benefits in gaining strength, and endurance exercise did not. Conclusion PAE induced a significant decrease in strength compared to CTRL in PND21. Resistance exercise is the most effective to reverse the effects of PAE on muscular strength. Our data suggests that individualized, scheduled, and supervised training of resistance is more beneficial than endurance or enriched environment exercise for adolescents FASD.

Methanotrophic bacteria are currently used industrially for the bioconversion of methane-rich natural gas and anaerobic digestion-derived biogas to valuable products. These bacteria may also serve to mitigate the negative effects of climate change by capturing atmospheric greenhouse gases. Several genetic tools have previously been developed for genetic and metabolic engineering of methanotrophs. However, the available tools for use in methanotrophs are significantly underdeveloped compared to many other industrially relevant bacteria, which hinders genetic and metabolic engineering of these biocatalysts. As such, expansion of the methanotroph genetic toolbox is needed to further our understanding of methanotrophy and develop biotechnologies that leverage these unique microbes for mitigation and conversion of methane to valuable products. Here, we determined the copy number of three broad-host-range plasmids in Methylococcus capsulatus Bath and Methylosinus trichosporium OB3b, representing phylogenetically diverse Gammaproteobacterial and Alphaproteobacterial methanotrophs, respectively. Further, we show that the commonly used synthetic Anderson series promoters are functional and exhibit similar relative activity in M. capsulatus and M. trichosporium OB3b, but the synthetic series had limited range. Thus, we mutagenized the native M. capsulatus particulate methane monooxygenase promoter and identified variants with activity that expand the activity range of synthetic, constitutive promoters functional not only in M. capsulatus, but also in Escherichia coli. Collectively, the tools developed here advance the methanotroph genetic engineering toolbox and represent additional synthetic genetic parts that may have broad applicability in Pseudomonadota bacteria.

Background. Preimplantation biopsy combines measurements of injury into a composite index to inform organ acceptance. The uncertainty in these measurements remains poorly characterized, raising concerns variability may contribute to inappropriate clinical decisions. Methods. We adopted a metrological approach to evaluate biopsy score reliability. Variability was assessed by performing repeat biopsies (n = 293) on discarded allografts (n = 16) using 3 methods (core, punch, and wedge). Uncertainty was quantified using a bootstrapping analysis. Observer effects were controlled by semi-blinded scoring, and the findings were validated by comparison with standard glass evaluation. Results. The surgical method strongly determined the size (core biopsy area 9.04 mm2, wedge 37.9 mm2) and, therefore, yield (glomerular yield r = 0.94, arterial r = 0.62) of each biopsy. Core biopsies yielded inadequate slides most frequently. Repeat biopsy of the same kidney led to marked variation in biopsy scores. In 10 of 16 cases, scores were contradictory, crossing at least 1 decision boundary (ie, to transplant or to discard). Bootstrapping demonstrated significant uncertainty associated with single-slide assessment; however, scores were similar for paired kidneys from the same donor. Conclusions. Our investigation highlights the risks of relying on single-slide assessment to quantify organ injury. Biopsy evaluation is subject to uncertainty, meaning each slide is better conceptualized as providing an estimate of the kidney’s condition rather than a definitive result. Pooling multiple assessments could improve the reliability of biopsy analysis, enhancing confidence. Where histological quantification is necessary, clinicians should seek to develop new protocols using more tissue and consider automated methods to assist pathologists in delivering analysis within clinical time frames.

IntroductionRenal transplant biopsies provide insights into graft health and support decision making. The current evidence on links between biopsy scores and transplant outcomes suggests there may be numerous factors affecting biopsy scores. Here we adopt measurement science approach to investigate the sources of uncertainty in biopsy assessment and suggest techniques to improve its robustness.MethodsHistological assessments, Remuzzi scores, biopsy processing and clinical variables are obtained from 144 repeat biopsies originating from 16 deceased-donor kidneys. We conducted sensitivity analysis to find the morphometric features with highest discriminating power and studied the dependencies of these features on biopsy and stain type. The analysis results formed a basis for recommendations on reducing the assessment variability.ResultsMost morphometric variables are influenced by the biopsy and stain types. The variables with the highest discriminatory power are sclerotic glomeruli counts, healthy glomeruli counts per unit area, percentages of interstitial fibrosis and tubular atrophy as well as diameter and lumen of the worst artery. A revised glomeruli adequacy score is proposed to improve the robustness of the glomeruli statistics, whereby a minimum of 104 µm2 of cortex tissue is recommended to keep type 1 and type 2 error probabilities below 0.15 and 0.2.DiscussionThe findings are transferable to several biopsy scoring systems. We hope that this work will help practitioners to understand the sources of statistical uncertainty and improve the utility of renal biopsy.

ABSTRACT Methanotrophic bacteria play a vital role in the biogeochemical carbon cycle due to their unique ability to use CH4 as a carbon and energy source. Evidence suggests that some methanotrophs, including Methylococcus capsulatus, can also use CO2 as a carbon source, making these bacteria promising candidates for developing biotechnologies targeting greenhouse gas capture and mitigation. However, a deeper understanding of the dual CH4 and CO2 metabolism is needed to guide methanotroph strain improvements and realize their industrial utility. In this study, we show that M. capsulatus expresses five carbonic anhydrase (CA) isoforms, one α-CA, one γ-CA, and three β-CAs, that play a role in its inorganic carbon metabolism and CO2-dependent growth. The CA isoforms are differentially expressed, and transcription of all isoform genes is induced in response to CO2 limitation. CA null mutant strains exhibited markedly impaired growth compared to an isogenic wild-type control, suggesting that the CA isoforms have independent, non-redundant roles in M. capsulatus metabolism and physiology. Overexpression of some, but not all, CA isoforms improved bacterial growth kinetics and decreased CO2 evolution from CH4-consuming cultures. Notably, we developed an engineered methanotrophic biocatalyst overexpressing the native α-CA and β-CA with a 2.5-fold improvement in the conversion of CH4 to biomass. Given that product yield is a significant cost driver of methanotroph-based bioprocesses, the engineered strain developed here could improve the economics of CH4 biocatalysis, including the production of single-cell protein from natural gas or anaerobic digestion-derived biogas.IMPORTANCEMethanotrophs transform CH4 into CO2 and multi-carbon compounds, so they play a critical role in the global carbon cycle and are of interest for biotechnology applications. Some methanotrophs, including Methylococcus capsulatus, can also use CO2 as a carbon source, but this dual one-carbon metabolism is incompletely understood. In this study, we show that M. capsulatus carbonic anhydrases are critical for this bacterium to optimally utilize CO2. We developed an engineered strain with improved CO2 utilization capacity that increased the overall carbon conversion to cell biomass. The improvements to methanotroph-based product yields observed here are expected to reduce costs associated with CH4 conversion bioprocesses.

Ultrastructural studies of contusive spinal cord injury (SCI) in mammals have shown that the most prominent acute changes in white matter are periaxonal swelling and separation of myelin away from their axon, axonal swelling, and axonal spheroid formation. However, the underlying cellular and molecular mechanisms that cause periaxonal swelling and the functional consequences are poorly understood. We hypothesized that periaxonal swelling and loss of connectivity between the axo-myelinic interface impedes neurological recovery by disrupting conduction velocity, and glial to axonal trophic support resulting in axonal swelling and spheroid formation. Utilizing in vivo longitudinal imaging of Thy1YFP+ axons and myelin labeled with Nile red, we reveal that periaxonal swelling significantly increases acutely following a contusive SCI (T13, 30 kdyn, IH Impactor) versus baseline recordings (laminectomy only) and often precedes axonal spheroid formation. In addition, using longitudinal imaging to determine the fate of myelinated fibers acutely after SCI, we show that ∼73% of myelinated fibers present with periaxonal swelling at 1 h post SCI and ∼ 51% of those fibers transition to axonal spheroids by 4 h post SCI. Next, we assessed whether cation-chloride cotransporters present within the internode contributed to periaxonal swelling and whether their modulation would increase white matter sparing and improve neurological recovery following a moderate contusive SCI (T9, 50 kdyn). Mechanistically, activation of the cation-chloride cotransporter KCC2 did not improve neurological recovery and acute axonal survival, but did improve chronic tissue sparing. In distinction, the NKKC1 antagonist bumetanide improved neurological recovery, tissue sparing, and axonal survival, in part through preventing periaxonal swelling and disruption of the axo-myelinic interface. Collectively, these data reveal a novel neuroprotective target to prevent periaxonal swelling and improve neurological recovery after SCI.

Background: Parkinson's disease (PD) is associated with both sleep disturbances and apathy, and within PD, apathy has been associated with REM behavior disorder and excessive daytime sleepiness. Whether other forms of sleep disturbance are similarly associated with apathy in PD remains unclear. This study explored associations between a broad array of sleep disturbances and apathy in 50 individuals with idiopathic PD (PD) and 48 matched controls (MC). Methods: Participants were adults aged 53–80 (Mdn = 67), 23 % female, and 96 % white. Sleep disturbances were measured with various questionnaires (ISI, PSQI, PROMIS-SD, ESS, PROMIS-SRI, RBDSQ). Mood was measured with the STAI and BDI-II. Apathy was evaluated using the Apathy Scale (AS). Spearman correlations and regression analyses were performed between measures of sleep disturbance and AS in the total sample and each group. Group correlations were compared using 2-sample Fisher's z test. Results: The AS total score significantly correlated with PROMIS-SRI in the total sample and multiple measures of sleep disturbance in the PD group. The apathy subscales were each significantly correlated with sleep disturbance measures in the total sample, MC, and PD groups. The correlations between several sleep and apathy values were significantly stronger in the PD group than MC. When accounting for anxiety and depression most differences were no longer significant, only the PROMIS-SRI was significantly predictive of the behavioral apathy sub score. Discussion: Evidence supports an association between sleep disturbances and apathy in individuals with PD. Specifically, insomnia severity, poor sleep quality, and daytime sleepiness were uniquely associated with apathy in this group. We did not find these associations in the matched control group. Anxiety and depression are likely involved in the association between sleep and apathy in PD. Experimental studies that manipulate or improve sleep may further elucidate the mechanisms underlying the association between sleep disturbance and apathy in PD.

PurposeDespite a growing number of women entering medical school, a small proportion of women pursue surgical specialties, including pediatric surgery. This multi-center study assesses gender-based differences in measures of scholarly productivity and distribution of faculty positions.MethodsThis is a retrospective web-based study of all pediatric surgeons at twelve large institutions across the United States. Data published by the American Association of Medical Colleges was compiled and analyzed to understand the gender distributions of medical students, general surgery residents, and pediatric surgery fellows. P-values were calculated using two-sided Student's independent t-tests and chi-squared tests.ResultsThere have been a growing number of women applying into pediatric surgery, but the proportion of women matriculating into these fellowships is not concordant. Women are still underrepresented (28%) amongst the pediatric surgeon workforce. A total of 111 pediatric surgeons were identified for this study, which included 31 women (28%) and 80 males (72%). There was a significant difference in the distribution across academic ranks between genders (p

Abstract Recreation is known to impact wildlife by displacing and sometimes contributing to the extirpation of sensitive species, underscoring a need for policies that balance wildlife and recreation. This is especially pressing when Indigenous rights necessitate ecological integrity and sustainable populations of wildlife throughout traditional territories. In the Cascade Mountain Range of Washington, USA, Indigenous harvest of elk (Cervus canadensis) is declining, concurrent with increases in recreation. Yet, the nature and magnitude of the effects of recreation on elk are unknown, which prevents land managers from developing informed policies regarding recreation and wildlife management. Here, we use camera traps alongside visitation models that incorporate geolocated social media to quantify the impacts of recreation on elk in western Washington. Random forest models show elk detection rates are relatively constant at low levels of recreation (0–11 people/day), but decrease by over 41% when recreation increases from 12 to 22 people/day. Activity overlap analysis also revealed a shift toward increased evening activity by elk at cameras with higher‐than‐average recreation (∆ = 0.70, 95% CI = 0.61–0.88; χ2 = 7.79, p = 0.02). Generalized additive modeling confirms that elk are more crepuscular or nocturnal at camera locations with more than 10 hiker detections per day. We compare methods for estimating recreation, showing model‐based estimates are more informative than camera‐based estimates. This indicates that recreational intensity along and in the immediate vicinity of trails may be a better predictor of impacts on wildlife than camera‐based estimates that quantify recreational intensity at specific locations along trails. We stress the importance of examining the impacts of recreation on wildlife across multiple spatiotemporal scales and underscore how novel approaches can provide land managers valuable tools to develop management strategies that balance recreation and wildlife. We hope that our work can also serve as a strong example of collaboration between universities, state agencies, and sovereign Indigenous nations toward the broader goal of mitigating the negative impacts of recreation on wildlife.

Abstract As the community increases climate model horizontal resolutions and experiments with removing moist convective parameterizations entirely, it is imperative to understand how these advances affect the InterTropical Convergence Zone (ITCZ). We investigate how the ITCZ responds to deactivating parameterized convection at two resolutions, 50 and 6 km, in fixed sea surface temperature, aquaplanet simulations with the NOAA GFDL AM4 atmospheric model. Disabling parameterized convection at 50 km resolution narrows the ITCZ and increases its precipitation minus evaporation (P–E) maximum by ∼78%, whereas at 6 km resolution doing so widens the ITCZ and decreases its P–E maximum by ∼50%. Using the column‐integrated moist static energy budget, we decompose these tropical P–E responses into contributions from changes in atmospheric energy input (AEI), gross moist stability, and gross moisture stratification. At 6 km, the ITCZ weakens due to increased gross moist stability. Disabling the convective parameterization at this finer resolution deepens the circulation, favoring more efficient poleward energy transport out of the deep tropics and reduced precipitation in the core of the ITCZ. Conversely, at 50 km the ITCZ strengthening is primarily driven by AEI, which in turn stems primarily from increased low cloud amount and thus longwave cloud radiative cooling in the Hadley cell subsiding branch. The Hadley circulation overturning intensifies to produce poleward energy fluxes that compensate the longwave cooling, yielding a stronger ITCZ. We further show that the low level diabatic heating profiles over the descending region are instrumental in understanding such diverse responses.

Vessel losses of command (e.g., propulsion, electrical power, or steering failure) can serve as the initiating event for significant maritime accidents, such as grounding. It is therefore important to estimate the frequency of such events for navigational risk assessments. Despite the existence of previous studies, new data have become available that may reveal more precise and useful information for risk estimations and accident prevention. This paper analyzes two databases of vessel losses of command within Norway’s economic exclusive area over a five year period. The study utilizes a novel database of incidents observed by the Norwegian Coastal Administration’s (NCA) Vessel Traffic Services (VTS), in addition to incidents reported to IHS Markit over the same five year period. Automatic identification system (AIS) data is used to construct activity metrics of vessel activity, which are used to compute vessel loss of command incidence rates for ship types. To account for under-reporting, capture–recapture methods are used to calculate adjusted incidence-rates. The duration of each incident is recorded and used to construct distributions of repair times. Probabilities of towing and anchoring are computed for ship types and incident location. The results indicate that previous studies may have underestimated the incidence of vessel loss of command. Using the best-case scenario, cargo ships and tankers suffer losses of command more frequently than other ship types. Significant under-reporting of losses of command was observed in the IHS Markit database.

Abstract Outdoor recreation is an essential way many people engage with nature. The provision of public spaces for recreation intersects with conservation practices motivated by intertwined social and ecological values, such as strict practices associated with the concept of ‘wilderness’. Debates persist about how such concepts and management practices influence people's recreation experiences. Many US public land management agencies facilitate opportunities for outdoor recreation, relying on management frameworks and tools intended to foster specific experiential qualities. But these frameworks and tools assume simplistic relationships between settings and people's experiences, and managers rarely assess these relationships. This study uses a data set of nearly 70,000 crowdsourced trip reports from a hiking website to understand the qualities of visitors' experiences on trails. We study the geographic distribution of experiential qualities commonly associated with US wilderness areas: aesthetics, awe, challenge, pristineness, quietness, solitude and timelessness. Using analytical methods that rely on machine learning and natural language processing, we identify these experiential qualities in trip reports from hundreds of routes, and use generalized linear models to analyse relationships between the frequency of each experiential quality and the route's administrative, built, biophysical, geographic and social settings. We find that four of the seven experiential qualities (aesthetics, awe, challenge and solitude) are commonly described in trip reports, each appearing in 15%–55% of manually coded reports. The extent to which setting characteristics explained variability in experiences differed, ranging from 34% of the variability in the proportion of trip reports describing aesthetics to 55% for awe. The setting characteristics associated with each experiential quality also differed, with characteristics such as trail mileage and summit destinations having stronger influences on experiential qualities than characteristics such as wilderness designation. Synthesis and applications. Our findings suggest the need to consider more diverse variables in experience–setting relationships, develop more robust models to characterize those relationships and create new data sources to represent understudied variables. These advances would help empirically inform and improve frameworks and tools used for recreation and wilderness planning and monitoring, and potentially promote more responsive management to evolving social–ecological values. Read the free Plain Language Summary for this article on the Journal blog.

Introduction/Purpose: Ankle fractures are one of the most common fractures encountered in orthopedics. The Maisonneuve fracture pattern is described as a pronation, external rotation injury involving the medial ankle structures, the syndesmosis, and the proximal third of the fibula. However, the actual distance from the proximal fibula has not been defined in what distinguishes a Maisonneuve fracture. With Weber C fractures, most surgeons tend to provide fixation to the fibula and reassess the syndesmosis afterwards. When dealing with the proximal fibula “Maisonneuve” fracture pattern, most surgeons would tend to ignore the fibula fracture and focus on syndesmotic reduction. Orthopaedic Trauma Association and American Orthopaedic Foot & Ankle Society provided their opinion via survey in treating increasing proximity of fibula fractures associated with unstable ankle fracture patterns. Methods: A survey was provided to eight OTA and AOFAS orthopedic surgeons. A powerpoint was provided to the surgeons that contained non-weightbearing injury films of eighteen patients. A questionnaire was provided giving two answer choices, address the fibular or syndesmosis primarily. The eighteen ankle fractures were selected based on increasing proximity of the fibula fracture, which ranged from 4.5cm to 32.3cm. The ankle fractures were grouped into four categories to include the Maisonneuve variant based on distance. These fracture radiographs were randomized in order to not influence the surgeon’s opinion during the study. The four groups were as follows: 1. 4.5cm – 7.4cm to include six ankle fracture radiographs 2. 8cm – 10.4cm to include four ankle fracture radiographs 3. 14.6cm to 23.3cm to include five ankle fracture radiographs 4. 30.7cm to 32.3 cm to include three Maisonneuve variant ankle fracture radiographs Results: Regarding section 1, the majority of surgeons responded with open reduction, internal fixation of the fibula as their initial reduction. Section 2, the responses remained consistent with a majority of surgeons choosing to address fibular fixation followed by syndesmotic evaluation. The total number of responses in this section scored 43 answer A selections to 5 answer B selections. Section 3 (14.6-23.3cm) provided the most variability in the responses provided. With 60 possible answer choices, the polled surgeons responded with answer choice A seventeen times and answer choice B 43 times. Section 4 (30.7-32.2cm) or the Maisonneuve produced a steady response of answer choice B. Syndesmotic reduction was performed 34 times compared to only two fibular fixation choices – or answer choice A. Conclusion: The purpose of this study was to evaluate expert opinion on differing treatment as the proximity of the fibular fracture increased in connection with an unstable ankle fracture pattern. General consensus under 10,4cm was to address fibular fixation. However, once the fibular fracture exceeded 14cm, significant variability was noted. These results prove that further biomechanical studies are needed to determine the effect fibular stability in increasing proximity has on the syndesmosis. Chart 1 shows answer selections across the four categories. Chart 2 shows answer selections across the highest variability group 3.

The ongoing phagocytic activity of macrophages necessitates an extraordinary capacity to digest and resolve incoming material. While the initial steps leading to the formation of a terminal phagolysosome are well studied, much less is known about the later stages of this process, namely the degradation and resolution of the phagolysosomal contents. We report that the degradation of targets such as splenocytes and erythrocytes by phagolysosomes occurs in a stepwise fashion, requiring lysis of their plasmalemmal bilayer as an essential initial step. This is achieved by the direct extraction of cholesterol facilitated by Niemann-Pick protein type C2 (NPC2), which in turn hands off cholesterol to NPC1 for export from the phagolysosome. The removal of cholesterol ulimately destabilizes and permeabilizes the membrane of the phagocytic target, allowing access of hydrolases to its internal compartments. In contrast, we found that saposins, which activate the hydrolysis of sphingolipids, are required for lysosomal tubulation, yet are dispensable for the resolution of targets by macrophages. The extraction of cholesterol by NPC2 is therefore envisaged as rate-limiting in the clearance of membrane-bound targets such as apoptotic cells. Selective cholesterol removal appears to be a primary mechanism that enables professional phagocytes to distinguish the target membrane from the phagolysosomal membrane and may be conserved in the resolution of autolysosomes.

Abstract The logistics and delivery industry is undergoing a technology-driven transformation, with robotics, drones, and autonomous vehicles expected to play a key role in meeting the growing challenges of last-mile delivery. To understand the public acceptability of automated parcel delivery options, this U.S. study explores customer preferences for four innovations: autonomous vehicles, aerial drones, sidewalk robots, and bipedal robots. We use an Integrated Nested Choice and Correlated Latent Variable (INCLV) model to reveal substitution effects among automated delivery modes in a sample of U.S. respondents. The study finds that acceptance of automated delivery modes is strongly tied to shipment price and time, underscoring the importance of careful planning and incentives to maximize the trialability of innovative logistics options. Older individuals and those with concerns about package handling exhibit a lower preference for automated modes, while individuals with higher education and technology affinity exhibit greater acceptance. These findings provide valuable insights for logistics companies and retailers looking to introduce automation technologies in their last-mile delivery operations, emphasizing the need to tailor marketing and communication strategies to meet customer preferences. Additionally, providing information about appropriate package handling by automated technologies may alleviate concerns and increase the acceptance of these modes among all customer groups.

Due to lack of apex predators in human-dominated landscapes, mesopredator relationships are complex and spatiotemporal niche partitioning strategies can vary, especially when seasonal shifts in resource availability occur. Our objective was to understand spatiotemporal niche overlap across seasons among mesopredators inhabiting a barrier island complex. We placed 19 unbaited cameras throughout Fort De Soto County Park, Florida, USA between February 2021 and July 2023. Of six mesopredator species detected, three species had >75 detections during both the wet and dry seasons (coyote, Canis latrans; Virginia opossum, Didelphis virginiana; and raccoon, Procyon lotor). Using general linear mixed models, we determined that during the wet season coyote–raccoon and raccoon–opossum detections were positively associated with each other (p < 0.05). During the dry season, raccoon–opossum detections were positively associated, and opossums were more likely to be detected around mangroves. After calculating coefficients of overlap, we found all three species varied their temporal activity between seasons. During the dry season exclusively, all three mesopredators occupied different temporal niches. The park’s isolated but developed nature has potentially led to a destabilized mesopredator community. Understanding seasonal mesopredator dynamics of Fort De Soto is particularly important because this park supports a high number of nesting shorebirds and sea turtles, which are known food sources for mesopredators.

Abstract Background Staphylococcus aureus can infect and adapt to multiple host species. However, our understanding of the genetic and evolutionary drivers of its generalist lifestyle remains inadequate. This is particularly important when considering local populations of S. aureus, where close physical proximity between bacterial lineages and between host species may facilitate frequent and repeated interactions between them. Here, we aim to elucidate the genomic differences between human- and animal-derived S. aureus from 437 isolates sampled from disease cases in the northeast region of the United States. Results Multi-locus sequence typing revealed the existence of 75 previously recognized sequence types (ST). Our population genomic analyses revealed heterogeneity in the accessory genome content of three dominant S. aureus lineages (ST5, ST8, ST30). Genes related to antimicrobial resistance, virulence, and plasmid types were differentially distributed among isolates according to host (human versus non-human) and among the three major STs. Across the entire population, we identified a total of 1,912 recombination events that occurred in 765 genes. The frequency and impact of homologous recombination were comparable between human- and animal-derived isolates. Low-frequency STs were major donors of recombined DNA, regardless of the identity of their host. The most frequently recombined genes (clfB, aroA, sraP) function in host infection and virulence, which were also frequently shared between the rare lineages. Conclusions Taken together, these results show that frequent but variable patterns of recombination among co-circulating S. aureus lineages, including the low-frequency lineages, that traverse host barriers shape the structure of local gene pool and the reservoir of host-associated genetic variants. Our study provides important insights to the genetic and evolutionary factors that contribute to the ability of S. aureus to colonize and cause disease in multiple host species. Our study highlights the importance of continuous surveillance of S. aureus circulating in different ecological host niches and the need to systematically sample from them. These findings will inform development of effective measures to control S. aureus colonization, infection, and transmission across the One Health continuum.

Introduction Worldwide, pancreatic cancer has a poor prognosis. Early diagnosis may improve survival by enabling curative treatment. Statistical and machine learning diagnostic prediction models using risk factors such as patient demographics and blood tests are being developed for clinical use to improve early diagnosis. One example is the Enriching New-onset Diabetes for Pancreatic Cancer (ENDPAC) model, which employs patients’ age, blood glucose and weight changes to provide pancreatic cancer risk scores. These values are routinely collected in primary care in the UK. Primary care’s central role in cancer diagnosis makes it an ideal setting to implement ENDPAC but it has yet to be used in clinical settings. This study aims to determine the feasibility of applying ENDPAC to data held by UK primary care practices.Methods and analysis This will be a multicentre observational study with a cohort design, determining the feasibility of applying ENDPAC in UK primary care. We will develop software to search, extract and process anonymised data from 20 primary care providers’ electronic patient record management systems on participants aged 50+ years, with a glycated haemoglobin (HbA1c) test result of ≥48 mmol/mol (6.5%) and no previous abnormal HbA1c results. Software to calculate ENDPAC scores will be developed, and descriptive statistics used to summarise the cohort’s demographics and assess data quality. Findings will inform the development of a future UK clinical trial to test ENDPAC’s effectiveness for the early detection of pancreatic cancer.Ethics and dissemination This project has been reviewed by the University of Surrey University Ethics Committee and received a favourable ethical opinion (FHMS 22-23151 EGA). Study findings will be presented at scientific meetings and published in international peer-reviewed journals. Participating primary care practices, clinical leads and policy makers will be provided with summaries of the findings.

The sustainability of marine communities is critical for supporting many biophysical processes that provide ecosystem services that promote human well-being. It is expected that anthropogenic disturbances such as climate change and human activities will tend to create less energetically-efficient ecosystems that support less biomass per unit energy flow. It is debated, however, whether this expected development should translate into bottom-heavy (with small basal species being the most abundant) or top-heavy communities (where more biomass is supported at higher trophic levels with species having larger body sizes). Here, we combine ecological theory and empirical data to demonstrate that full marine protection promotes shifts towards top-heavy energetically-efficient structures in marine communities. First, we use metabolic scaling theory to show that protected communities are expected to display stronger top-heavy structures than disturbed communities. Similarly, we show theoretically that communities with high energy transfer efficiency display stronger top-heavy structures than communities with low transfer efficiency. Next, we use empirical structures observed within fully protected marine areas compared to disturbed areas that vary in stress from thermal events and adjacent human activity. Using a nonparametric causal-inference analysis, we find a strong, positive, causal effect between full marine protection and stronger top-heavy structures. Our work corroborates ecological theory on community development and provides a quantitative framework to study the potential restorative effects of different candidate strategies on protected areas.

Public land managers depend on reliable and readily available data about outdoor recreation in parks and greenspaces. However, traditional recreation monitoring techniques including visitor surveying and counting cannot be implemented over large spatial and temporal scales, especially in remote and undeveloped settings where monitoring is costly. To fill these data gaps, and thereby inform decision-making, this study develops and tests the efficacy of a novel recreation monitoring technique that engages visitors in data collection using a chatbot and text-messages. Drawing on knowledge and methods from community science and crowdsourcing, we present a relatively low-cost and low-barrier approach to counting and characterizing recreational visits on public lands. In an 18-month pilot implementation on a national forest in Washington, USA, we found that crowdsourced data collected using the chatbot were consistent with results of controlled counts and in-person surveys. Furthermore, some sites received relatively high participation rates, up to 12% of recreating parties, regardless of cellular connectivity at the site. This study, which is the first to engage public land usersin community science using a text-messaging chatbot for the purposes of studying outdoor recreation, demonstrates the potential for technology to support new community science approaches that involve visitors in land stewardship and the development of recreation monitoring systems.

Abstract An idealized aquaplanet moist global atmospheric model with realistic radiative transfer but no clouds and no convective parameterization is found to possess multiple climate equilibria. When forced symmetrically about the equator, in some cases the Inter Tropical Convergence Zone (ITCZ) migrates to an off‐equatorial equilibrium position. Mechanism denial experiments prescribing relative humidity imply that radiation‐circulation coupling is essential to this instability. The cross‐equatorial asymmetry occurs only when the underlying slab ocean is sufficiently deep and the atmosphere's spectral dynamical core is sufficiently coarse (∼T170 or less with our control parameters). At higher resolutions, initializing with an asymmetric state indicates metastability with very slow (thousands of days) return to hemispheric symmetry. There is some sensitivity to the model timestep, which affects the time required to transition to the asymmetric state, with little effect on the equilibrium climate. The instability is enhanced when the planetary boundary layer scheme favors deeper layers or by a prescribed meridional heat transport away from the equator within the slab. The instability is not present when the model is run with a convective parameterization scheme commonly utilized in idealized moist models. We argue that the instability occurs when the asymmetric heating associated with a spontaneous ITCZ shift drives a circulation that rises poleward of the perturbed ITCZ. These results serve as a warning of the potential for instability and non‐uniqueness of climate that may complicate studies with idealized models of the tropical response to perturbations in forcing.

[INLINE:1] The formation of axonal spheroid is a common feature following spinal cord injury. To further understand the source of Ca2+ that mediates axonal spheroid formation, we used our previously characterized ex vivo mouse spinal cord model that allows precise perturbation of extracellular Ca2+. We performed two-photon excitation imaging of spinal cords isolated from Thy1YFP+ transgenic mice and applied the lipophilic dye, Nile red, to record dynamic changes in dorsal column axons and their myelin sheaths respectively. We selectively released Ca2+ from internal stores using the Ca2+ ionophore ionomycin in the presence or absence of external Ca2+. We reported that ionomycin dose-dependently induces pathological changes in myelin and pronounced axonal spheroid formation in the presence of normal 2 mM Ca2+ artificial cerebrospinal fluid. In contrast, removal of external Ca2+ significantly decreased ionomycin-induced myelin and axonal spheroid formation at 2 hours but not at 1 hour after treatment. Using mice that express a neuron-specific Ca2+ indicator in spinal cord axons, we confirmed that ionomycin induced significant increases in intra-axonal Ca2+, but not in the absence of external Ca2+. Periaxonal swelling and the resultant disruption in the axo-myelinic interface often precedes and is negatively correlated with axonal spheroid formation. Pretreatment with YM58483 (500 nM), a well-established blocker of store-operated Ca2+ entry, significantly decreased myelin injury and axonal spheroid formation. Collectively, these data reveal that ionomycin-induced depletion of internal Ca2+ stores and subsequent external Ca2+ entry through store-operated Ca2+ entry contributes to pathological changes in myelin and axonal spheroid formation, providing new targets to protect central myelinated fibers.

Abstract Escherichia coli (E. coli) are typically present as commensal bacteria in the gastro-intestinal tract of most animals including poultry species, but some avian pathogenic E. coli (APEC) strains can cause localized and even systematic infections in domestic poultry. Emergence and re-emergence of antimicrobial resistant isolates (AMR) constrain antibiotics usage in poultry production, and development of an effective vaccination program remains one of the primary options in E. coli disease prevention and control for domestic poultry. Thus, understanding genetic and pathogenic diversity of the enzootic E. coli isolates, particularly APEC, in poultry farms is the key to designing an optimal vaccine candidate and to developing an effective vaccination program. This study explored the genomic and pathogenic diversity among E. coli isolates in southern United States poultry. A total of nine isolates were recovered from sick broilers from Mississippi, and one from Georgia, with epidemiological variations among clinical signs, type of housing, and bird age. The genomes of these isolates were sequenced by using both Illumina short-reads and Oxford Nanopore long-reads, and our comparative analyses suggested data from both platforms were highly consistent. The 16 s rRNA based phylogenetic analyses showed that the 10 bacteria strains are genetically closer to each other than those in the public database. However, whole genome analyses showed that these 10 isolates encoded a diverse set of reported virulence and AMR genes, belonging to at least nine O:H serotypes, and are genetically clustered with at least five different groups of E. coli isolates reported by other states in the United States. Despite the small sample size, this study suggested that there was a large extent of genomic and serological diversity among E. coli isolates in southern United States poultry. A large-scale comprehensive study is needed to understand the overall genomic diversity and the associated virulence, and such a study will be important to develop a broadly protective E. coli vaccine.

Neuronal ryanodine receptors (RyR) release calcium from internal stores and play a key role in synaptic plasticity, learning, and memory. Dysregulation of RyR function contributes to neurodegeneration and negatively impacts neurological recovery after spinal cord injury (SCI). However, the individual role of RyR isoforms and the underlying mechanisms remain poorly understood. To determine whether RyR2 plays a direct role in axonal fate and functional recovery after SCI, we bred Advillin-Cre: tdTomato (Ai9) reporter mice with ?floxed? RyR2 mice to directly knock out (KO) RyR2 function in dorsal root ganglion neurons and their spinal projections. Adult 6- to 8-week-old RyR2KO and littermate controls were subjected to a contusive SCI and their dorsal column axons were imaged in vivo using two-photon excitation microscopy. We found that direct RyR2KO in dorsal column primary afferents did not significantly alter secondary axonal degeneration after SCI. We next assessed behavioral recovery after SCI and found that direct RyR2KO in primary afferents worsened open-field locomotor scores (Basso Mouse Scale subscore) compared to littermate controls. However, both TreadScan? gait analysis and overground kinematic gait analysis tests revealed subtle, but no fundamental, differences in gait patterns between the two groups after SCI. Subsequent removal of spared afferent fibers using a dorsal column crush revealed similar outcomes in both groups. Analysis of primary afferents at the lumbar (L3?L5) level similarly revealed no noticeable differences between groups. Together, our results support a modest contribution of dorsal column primary afferent RyR2 in neurological recovery after SCI.

Abstract The Bowfin (Amia calva), as currently recognized, represents the sole living member of the family Amiidae, which dates back to approximately 150 Ma. Prior to 1896, 13 species of extant Bowfins had been described, but these were all placed into a single species with no rationale or analysis given. This situation has persisted until the present day, with little attention given to re-evaluation of those previously described nominal forms. Here, we present a phylogenomic analysis based on over 21,000 single nucleotide polymorphisms (SNPs) from 94 individuals that unambiguously demonstrates the presence of at least two independent evolutionary lineages within extant Amia populations that merit species-level standing, as well as the possibility of two more. These findings not only expand the recognizable species diversity in an iconic, ancient lineage, but also demonstrate the utility of such methods in addressing previously intractable questions of molecular systematics and phylogeography in slowly evolving groups of ancient fishes.

Abstract Environmental management increasingly relies on information about ecosystem services for decision‐making. Compared with regulating and provisioning services, cultural ecosystem services (CES) are particularly challenging to characterize and measure at management‐relevant spatial scales, which has hindered their consideration in practice. Social media are one source of spatially explicit data on where environments support various types of CES, including physical activity. As tools for automating social media content analysis with artificial intelligence (AI) become more commonplace, studies are promoting the potential for AI and social media to provide new insights into CES. Few studies, however, have evaluated what biases are inherent to this approach and whether it is truly reproducible. This study introduces and applies a novel and open‐source convolutional neural network model that uses computer vision to recognize recreational activities in the content of photographs shared as social media. We train a model to recognize 12 common recreational activities to map one aspect of recreation in a national forest in Washington, USA, based on images uploaded to Flickr. The image classifier performs well, overall, but varies by activity type. The model, which is trained with data from one region, performs nearly as well in a novel region of the same national forest, suggesting that it is broadly applicable across similar public lands. By comparing the results from our CNN model with an on‐site survey, we find that there are apparent biases in which activities visitors choose to photograph and post to social media. After considering potential issues with underlying data and models, we map activity diversity and find that natural features (such as rivers, lakes and higher elevations) and some built infrastructure (campgrounds, trails, roads) support a greater diversity of activities in this region. We make our model and training weights available in open‐source software, to facilitate reproducibility and further model development by researchers who seek to understand recreational values at management‐relevant scales—and more broadly provide an example of how to build, test and apply AI to understand recreation and other types of CESs. Read the free Plain Language Summary for this article on the Journal blog.

Digital pathology is revolutionising the analysis of histological features and is becoming more and more widespread in both the clinic and research. Molecular pathology extends the tissue morphology information provided by conventional histopathology by providing spatially resolved molecular information to complement the structural information provided by histopathology. The multidimensional nature of the molecular data poses significant challenge for data processing, mining, and analysis. One of the key challenges faced by new and existing pathology practitioners is how to choose the most suitable molecular pathology technique for a given diagnosis. By providing a comparison of different methods, this narrative review aims to introduce the field of molecular pathology, providing a high-level overview of many different methods. Since each pixel of an image contains a wealth of molecular information, data processing in molecular pathology is more complex. The key data processing steps and variables, and their effect on the data, are also discussed.

Multi-scale instabilities are ubiquitous in atmospheric and oceanic flows and are essential topics in teaching geophysical fluid dynamics. Yet these topics are often difficult to teach and counter-intuitive to new learners. In this paper, we introduce our state-of-the-art Do-It Yourself Dynamics (DIYnamics) LEGO® robotics kit that allows users to create table-top models of geophysical flows. Deep ocean convection processes are simulated via three experiments – upright convection, thermal wind flows, and baroclinic instability – in order to demonstrate the robust multi-scale modeling capabilities of our kit. Detailed recipes are provided to allow users to reproduce these experiments. Further, dye-visualization measurements show that the table-top experimental results adequately agree with theory. In sum, our DIYnamics setup provides students and educators with an accessible table-top framework by which to model the multi-scale behaviors, inherent in canonical geophysical flows, such as deep ocean convection.

We present a uniform forward-modeling analysis of 90 late-M and L dwarfs in nearby young (∼10–200 Myr) moving groups, the Pleiades, and the Hyades using low-resolution ( R ≈ 150) near-infrared (0.9–2.4 μ m) spectra and the BT-Settl model atmospheres. We derive the objects’ effective temperatures, surface gravities, radii, and masses by comparing our spectra to the models using a Bayesian framework with nested sampling and calculate the same parameters using evolutionary models. Assuming the evolutionary-based parameters are more robust, our spectroscopically inferred parameters from BT-Settl exhibit two types of systematic behavior for objects near the M-L spectral type boundary. Several objects are clustered around T _eff ≈ 1800 K and $\mathrm{log}g\approx 5.5$ dex, implying impossibly large masses (150–1400 M _Jup ), while others are clustered around T _eff ≳ 3000 K and $\mathrm{log}g\lesssim 3.0$ dex, implying unphysically low masses and unreasonably young ages. We find the fitted BT-Settl model spectra tend to overpredict the peak J - and H -band flux for objects located near the M-L boundary, suggesting the dust content included in the model atmospheres is insufficient to match the observations. By adding an interstellar medium–like reddening law to the BT-Settl model spectra, we find the fits between models and observed spectra are greatly improved, with the largest reddening coefficients occurring at the M-L transition. This work delivers a systematic examination of the BT-Settl model atmospheres and constitutes the largest spectral analysis of benchmark late-M- and L-type brown dwarfs to date.

Abstract Necrosis seen in histopathology Whole Slide Images is a major criterion that contributes towards scoring tumour grade which then determines treatment options. However conventional manual assessment suffers from inter-operator reproducibility impacting grading precision. To address this, automatic necrosis detection using AI may be used to assess necrosis for final scoring that contributes towards the final clinical grade. Using deep learning AI, we describe a novel approach for automating necrosis detection in Whole Slide Images, tested on a canine Soft Tissue Sarcoma (cSTS) data set consisting of canine Perivascular Wall Tumours (cPWTs). A patch-based deep learning approach was developed where different variations of training a DenseNet-161 Convolutional Neural Network architecture were investigated as well as a stacking ensemble. An optimised DenseNet-161 with post-processing produced a hold-out test F1-score of 0.708 demonstrating state-of-the-art performance. This represents a novel first-time automated necrosis detection method in the cSTS domain as well specifically in detecting necrosis in cPWTs demonstrating a significant step forward in reproducible and reliable necrosis assessment for improving the precision of tumour grading.

Abstract Microbes in marine sediments constitute a large percentage of the global marine ecosystem and function to maintain a healthy food web. In continental shelf habitats such as the Gulf of Maine (GoM), relatively little is known of the microbial community abundance, biodiversity, and natural product potential. This report is the first to provide a time-series assessment (2017–2020) of the sediment microbial structure in areas open and closed to fishing within the Stellwagen Bank National Marine Sanctuary (SBNMS). A whole metagenome sequencing (WMS) approach was used to characterize the sediment microbial community. Taxonomic abundance was calculated across seven geographic sites with 14 individual sediment samples collected during the summer and fall seasons. Bioinformatics analyses identified more than 5900 different species across multiple years. Non-metric multidimensional scaling methods and generalized linear models demonstrated that species richness was inversely associated with fishing exposure levels and varied by year. Additionally, the discovery of 12 unique biosynthetic gene clusters (BGCs) collected across sites confirmed the potential for medically relevant natural product discovery in the SBNMS. This study provides a practical assessment of how fishing exposure and temporal trends may affect microbial community structure in a coastal marine sanctuary.

Abstract Using a dataset of more than 51,000 US lakes, we estimated the relationship between summertime lake visits, lake water clarity, landscape features, and other amenities, where visits were estimated with counts of geo‐located photographs. Given the size and complexity of our dataset, we used a combination of machine learning techniques, imputation techniques, and a Poisson count model to estimate these relationships. We found that every additional meter of average summertime Secchi depth was associated with at least 7% more summertime lake visits, all else equal. Second, we found that lake amenities, such as beaches, boat launches, and public toilets, were more powerful predictors of visits than water clarity. Third, we found that visits to a lake were strongly influenced by the lake's accessibility and its distance to nearby lakes and the amenities the nearby lakes offered. Our research highlights the need for (1) a better understanding of how representative social media data are of actual recreational behavior, (2) the development of best practices to account for nonrandom patterns in missing natural feature data, and (3) a better understanding of the potential endogeneity in the lake visit–water quality relationships.

Abstract The emergence of methicillin-resistant Staphylococcus aureus (MRSA) poses an important threat in human and animal health. In this study, we ask whether resistance and virulence genes in S. aureus are homogeneously distributed or constrained by different animal hosts. We carried out whole genome sequencing of 114 S. aureus isolates from ten species of animals sampled from four New England states (USA) in 2017–2019. The majority of the isolates came from cats, cows and dogs. The maximum likelihood phylogenetic tree based on the alignment of 89,143 single nucleotide polymorphisms of 1173 core genes reveal 31 sequence types (STs). The most common STs were ST5, ST8, ST30, ST133 and ST2187. Every genome carried at least eight acquired resistance genes. Genes related to resistance found in all genomes included norA (fluoroquinolone), arlRS (fluoroquinolone), lmrS (multidrug), tet(38) (tetracycline) and mepAR (multidrug and tigecycline resistance). The most common superantigen genes were tsst-1, sea and sec. Acquired antibiotic resistance (n = 10) and superantigen (n = 9) genes of S. aureus were widely shared between S. aureus lineages and between strains from different animal hosts. These analyses provide insights for considering bacterial gene sharing when developing strategies to combat the emergence of high-risk clones in animals.

BackgroundCalcineurin inhibitors (CNIs) are associated with nephrotoxicity, endothelial cell dysfunction, and thrombotic microangiopathy (TMA). Evolving evidence suggests an important role for complement dysregulation in the pathogenesis of CNI-induced TMA. However, the exact mechanism(s) of CNI-induced TMA remain(s) unknown.MethodsUsing blood outgrowth endothelial cells (BOECs) from healthy donors, we evaluated the effects of cyclosporine on endothelial cell integrity. Specifically, we determined complement activation (C3c and C9) and regulation (CD46, CD55, CD59, and complement factor H [CFH] deposition) as these occurred on the endothelial cell surface membrane and glycocalyx.ResultsWe found that exposing the endothelium to cyclosporine resulted in a dose- and time-dependent enhancement of complement deposition and cytotoxicity. We, therefore, employed flow cytometry, Western blotting/CFH cofactor assays, and immunofluorescence imaging to determine the expression of complement regulators and the functional activity and localization of CFH. Notably, while cyclosporine led to the upregulation of complement regulators CD46, CD55, and CD59 on the endothelial cell surface, it also diminished the endothelial cell glycocalyx through the shedding of heparan sulfate side chains. The weakened endothelial cell glycocalyx resulted in decreased CFH surface binding and surface cofactor activity.ConclusionOur findings confirm a role for complement in cyclosporine-induced endothelial injury and suggest that decreased glycocalyx density, induced by cyclosporine, is a mechanism that leads to complement alternative pathway dysregulation via decreased CFH surface binding and cofactor activity. This mechanism may apply to other secondary TMAs—in which a role for complement has so far not been recognized—and provide a potential therapeutic target and an important marker for patients on calcineurin inhibitors.

Abstract Marine cloud brightening is a proposal to counteract global warming by increasing sea salt aerosol emissions. In theory, this increases the cloud droplet number concentration of subtropical marine stratocumulus decks, increasing cloud brightness and longevity. However, this theoretical progression remains uncertain in coupled climate models, especially the response of liquid water path and cloud fraction to aerosol seeding. We use the GFDL CM4 climate model to simulate marine cloud brightening following the published G4sea‐salt protocol, in which sea salt aerosol emissions are uniformly increased over 30 S–30 N in addition to standard forcings from a SSP2‐4.5 future warming scenario. The perturbed radiative and cloud responses are temporally stable though spatially heterogeneous, and direct scattering by the added sea salt predominates over changes to cloud reflectance. In fact, feedbacks in the coupled simulation lead to a net warming, rather than cooling, response by clouds.